The objectives of this grant are to study collagen and fibrous protein metabolism in heritable disorders of connective tissue and acquired diseases, such as scleroderma, to attempt to elucidate principles of normal collagen biochemistry, and to develop methods of studying collagen metabolism in human diseases. Several projects are currently underway which will be continued through the coming year. The purification and characterization of basement membranes collagens has proved elusive because of the lack of a suitable system for obtaining pure basement membrane and the inability to solubilize the collagenous components. We are attempting to characterize these collagens using limited protease digestion of swine anterior lens capsule and differential salt fractionation of the solubilized collagen. Preliminary results indicate that three distinct collagenous species are isolated by this procedure. Fibronectin, the major cell surface protein, has also been identified as a collagen binding protein which probably functions to attatch fibroblasts to the collagen substratum. We have purified fibronectin, prepared a specific antibody to it and are attempting to assess its role in connective tissue disease by assessing blood levels by radioimmunoassay. Antibodies to fibroblast cell surface proteins, such as antibody to Beta-2 microglobulins induce complement mediated cytotoxicity (CMC). Purified bacterial collagenase, but not proteolytic enzymes, abrogate this CMC. We are attempting to ascertain if this phenomenon occurs because bacterial collagenase destroys a specific fibroblast complement receptor which is collagenase sensitive. Over the past year, we have ascertained 10 families with the Ehlers-Danlos Syndrome (EDS) through the arthritis clinic. These patients have presented with recurrent arthralgias and joint swelling with no diagnostic radiologic or serologic manifestations. Most of these patients have had EDS type II or type III. Over the coming year, these patients will be utilized to study collagen metabolism in EDS.